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Method for extracting and purifying saxitoxin from poisonous alga

A shellfish toxin and algae technology, applied in chemical instruments and methods, separation methods, preparation of test samples, etc., can solve the problems of low content of shellfish toxins, unsatisfactory yields, cumbersome steps, etc., to save manpower and material resources, Improved yield and purity, simple effect of extraction and purification steps

Inactive Publication Date: 2018-04-17
上海泰坦科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the content of shellfish toxin obtained by the current extraction and purification method is low, the purity is not high, and the steps are cumbersome, and the yield is not satisfactory

Method used

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  • Method for extracting and purifying saxitoxin from poisonous alga
  • Method for extracting and purifying saxitoxin from poisonous alga

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] A method for extracting and purifying shellfish toxins from toxin-producing algae, comprising the steps of:

[0042] (1) Extraction: add toxin-producing algae into 2L natural seawater culture medium according to the wet weight of 50g, and centrifuge at a speed of 5000rpm after cultivating for 48 hours. The mixed solution of chloroform and methanol was soaked for 30min, the volume ratio of chloroform and methanol was 1:1, and then the cells were disrupted by ultrasound with an intensity of 20%, and evaporated under reduced pressure to obtain the total extract;

[0043] (2) Extraction: add deionized water equivalent to 3 times the volume of the total extract to the total extract to dissolve it, use an extractant 3 times the volume of the total extract, add the first extractant, and mix thoroughly Stand still for 20 minutes, extract until the color is light and then change to another one, follow this step to add n-hexane, chloroform, ethyl acetate and n-butanol in sequence...

Embodiment 2

[0046] A method for extracting and purifying shellfish toxins from toxin-producing algae, comprising the steps of:

[0047] (1) Extraction: add toxin-producing algae into 2L natural seawater culture medium according to the wet weight of 50g, and centrifuge at a speed of 2000rpm after culturing for 48 hours. The mixed solution of chloroform and methanol was soaked for 30min, and the volume ratio of chloroform and methanol was 10:1, and then the cells were disrupted by ultrasound with an intensity of 10%, and evaporated under reduced pressure to obtain the total extract;

[0048] (2) Extraction: add deionized water equivalent to 2 times the volume of the total extract to the total extract to dissolve it, use an extractant twice the volume of the total extract, add the first extractant, and mix thoroughly Stand still for 20 minutes, extract until the color is light and then change to another one, follow this step to add n-hexane, chloroform, ethyl acetate and n-butanol in sequenc...

Embodiment 3

[0051] A method for extracting and purifying shellfish toxins from toxin-producing algae, comprising the steps of:

[0052] (1) Extraction: add toxin-producing algae into 2L natural seawater culture solution according to the wet weight of 50g, and centrifuge at a speed of 6000rpm after cultivating for 48 hours. The mixed solution of chloroform and methanol was soaked for 20min, and the volume ratio of chloroform and methanol was 8:1, and then the cells were disrupted by ultrasound with an intensity of 15%, and evaporated under reduced pressure to obtain the total extract;

[0053] (2) Extraction: add deionized water equivalent to 2 times the volume of the total extract to the total extract to dissolve it, use an extractant that is 4 times the volume of the total extract, add the first extractant, and mix thoroughly Stand still for 20 minutes, extract until the color is light and then change to another one, follow this step to add n-hexane, chloroform, ethyl acetate and n-butan...

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Abstract

The invention relates to a method for extracting and purifying saxitoxin from a poisonous alga. The method mainly comprises three steps of extraction, leaching and column chromatography. Extraction liquid comprises methyl alcohol and chloroform, a leaching agent comprises normal hexane, chloroform, ethyl acetate and normal butanol, and column chromatography comprises a positive and negative silicagel column layer and a high performance liquid chromatography. By optimizing the technological process, an eluent, the extraction liquid and the leaching agent are reasonably selected, through synergistic interaction of the steps, the saxitoxin which is high in purity and yield is finally obtained, the yield can reach 4.58 g, and the purity reaches 99%. The method is simple and efficient, the costis saved, the yield is relatively high, and the method is beneficial for industrial large-scale application.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a method for extracting and purifying shellfish toxins from toxin-producing algae. Background technique [0002] Marine toxins are toxic polymer compounds formed by the toxic algae in the ocean passing through the food chain to algae-eating fish, shrimp and shellfish, and accumulating in their bodies. There are many types of marine toxins, among which shellfish toxins are one of the most harmful. Shellfish toxins include paralytic shellfish toxins, diarrheal shellfish toxins, neurogenic shellfish toxins, and amnestic shellfish toxins. The hazards of shellfish toxins are sudden and widespread, and have the characteristics of strong toxicity and unpredictability. Due to their high toxicity, fast response, and no suitable antidote, many difficulties have been brought to the prevention and control. Shellfish toxins are harmful to the health of consumers. The serious threat to safety and c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N30/06B01D15/22
Inventor 谢应波张庆张华徐肖冰罗桂云张维燕
Owner 上海泰坦科技股份有限公司
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